5.5.1. Sample preparation for analysis by flow cytometry
After seeding the cells and inducing cell death as described in
Section 3.2.2, add TMRM to the cells (30–200 nM final). Cell samples
from suspension cultures are analyzed at regular intervals
on the flow cytometer. For this, 270 ll of cells from the 24-well
suspension plate is transferred to a 5-ml polypropylene round bottom
tube, and 30 ll of cell-impermeant probe, e.g. PI, is added from
a 10 solution (30 lM). A synchronic cellular response in DWm results
in an overall increase or decrease of the mean fluorescence
detected, while an asynchronic response is detected by a change
in the amount of cells showing increased or decreased DWm.